SINGLE AND PARALLEL DYE-BASED REAL-TIME PCR DETECTION OF FOODBORNE PATHOGENS SALMONELLA ENTERICA AND CAMPYLOBACTER JEJUNI.

Salmonella enterica and Campylobacter jejuni are some of the common foodborne pathogens causing gastrointestinal illnesses worldwide. The development of sensitive and specific detection methods is essential to ensure food safety. Dye-based real-time PCR assay using SYBR™ GreenER™ dye was developed for the detection of Salmonella enterica and Campylobacter jejuni. Designed primer sets specifically targeting the genes ompF and omp50 in Salmonella and Campylobacter, respectively, were utilised in the study. The assay was able to detect Salmonella and Campylobacter at as low as 50 fg/μl and 10 fg/μl, respectively. Specificity analysis performed using 16 different bacterial strains to check for cross- reactivity with the respective bacteria found the assay to be specific to Salmonella and Campylobacter. The assay successfully detected Salmonella enterica in inoculated food at as low as 5 fg/reaction for some food samples. Meanwhile, the detection limit for Campylobacter jejuni in all inoculated food samples was 2000 fg/reaction. The coefficient variations (CV%) of the assays for both pathogens indicated that the assays were highly reproducible. Therefore, the developed real-time PCR assays for both Salmonella enterica and Campylobacter jejuni detections were specific and sensitive and can be used for rapid screening to detect these foodborne pathogens. [ABSTRACT FROM AUTHOR]